Influence of microwave irradiation on the intraparticle diffusion of an insulin variant in reversed-phase liquid chromatography under linear conditions

Abstract

The influence of microwave irradiation on the mass transfer kinetics of an insulin variant in reversed-phase liquid chromatography (RPLC) was investigated. The elution band profiles of insulin were obtained by the pulse-response method, under linear conditions. The RPLC column was placed in a microwave oven and the incremental change in the temperature of the column effluent stream at various microwave energies and mobile phase flow rates were measured. The microwave energy dissipated in the column was set at 15 and 30 W and the mobile phase flow rate was varied from 1.0 to 2.5 mL/min at a mobile phase composition of acetonitrile, water, and trifloroacetic acid (31:69:0.1, v/v/v). The experimental data were analyzed using the conventional method of moment analysis and the lumped pore diffusion model. Regardless of mobile flow rates, the effluent temperatures measured at 15 and 30 W microwave power input were 25 ± 1 and 30 ± 1 °C, respectively. The effect of microwave irradiation on the mass transfer of the variant insulin was determined by comparing the band profiles obtained under the same experimental conditions, at the same column temperature, with and without irradiation. The calculated intraparticle diffusion coefficient, D e, at 30 W (30 ± 1 °C) microwave irradiation was ca. 20% higher than without irradiation at 30 ± 1 °C. These preliminary results suggest that microwave irradiation may have a significant influence on the intraparticle diffusion of insulin in RPLC.